The present invention relates to a novel 3-hydroxychromen-4-one derivative represented by the following formula (1): 
in which
A represents hydrogen or nitro, or represents amino which is optionally substituted by C1-C4-alkylcarbonyl or carbamoyl, or represents a structure selected from a group consisting of 
wherein R4 represents hydrogen or C1-C6-alkyl which is optionally substituted by amino or hydroxy, R3 represents C1-C6-alkyl which is optionally substituted by amino or hydroxy, and D represents halogen,
B represents methyl, or represents amino which is optionally mono- or disubstituted by substituents selected from a group consisting of C1-C6-alkyl, hydroxy-C1-C6-alkyl, C3-C6-cycloalkyl, acetyl, phenyl, benzyl and piperidinyl,
X, Y and Z independently of one another represent hydrogen, hydroxy, nitro, cyano or halogen, or represent amino which is optionally substituted by C1-C4-alkyl, C1-C4-alkylcarbonyl or carbamoyl, or represent C1-C4-alkyl which is optionally substituted by hydroxy or halogen,
pharmaceutically acceptable salt, hydrate, solvate or isomer thereof which is useful as an inhibitor for Cyclin Dependent Kinase (hereinafter, referred to as xe2x80x9cCDKxe2x80x9d).
The present invention further relates to a process for preparing the compound of formula (1) and to a composition for suppression or treatment of cancer and diseases induced by cell proliferation such as inflammation, angiostenosis, angiogenesis, etc. which comprises the compound of formula (1) as an active component together with pharmaceutically acceptable carriers.
Researches on cell division process in molecular level have been extensively performed from the late 1980""s through study of division of frog oocytes, analysis several yeast cell growth or characterization of induced mutants by radiation and study of the tumor suppressor Rb. In the 1990""s, it was discovered that cell growth regulators of small size control the cell division process (i.e. growth, differentiation, cytogenesis, aging and apoptosis, etc.) through their own regulatory function. These results were very useful for more precise understanding of the pathology of several diseases. A representative example is cancer. In transformation process from normal cells to cancer cells, it was frequently observed that cell growth regulators lose their own function. That is, in cancer cells, the cell growth regulators show an abnormal activity, which is intimately associated with invasion/metastasis, the most crucial factor considered in the cancerpathology. Particularly, cell cycle deregulation is recognized to be a direct cause of cancer since cancer occurs when overexpression or knock-out of cell growth regulators is induced in the transformed animals.
The cell growth is under positive or negative regulation in the same manner as other biological regulations. The major pathway of cell cycle regulation known up to now is based on CDK activity and as a result of studies on many cancer cells and carcinogenesis mechanisms, it was confirmed that problems of positive or negative regulation on CDK activity result in carcinogenesis in many cases. That is, cancer may occur when well-balanced regulation or timely regulation is upset.
The representative CDKs of mammals are CDK4(cyclin dependent kinase 4) which shows its activity in mid-G1 phase of cell cycle, CDK2 which shows its activity in mid-G1 and S phases, CDC2(CDK1) which shows its activity in G2-M phase. It has been known that CDK4 and CDK2 activities are regulated by check point of G1-S cell cycle and CDC2 activity by check point of G2-M. In many cancer cells, abnormalities appear in the regulatory mechanism of CDK4, CDK2 and CDC2(CDK1) and in fact, it was confirmed that artificially induced abnormalities cause cancer in the transformed animals. Therefore, the typical cyclin dependent kinases, i.e., CDK4, CDK2 and CDC2(CDK1) are suitable as a target of anti-cancer agents. Those kinases also become a target in developing an agent for suppression or treatment of cancer and diseases induced by cell proliferation such as inflammation, angiostenosis, angiogenesis, etc.
The results of studies on the relation between these CDKs and carcinogenesis will be explained in more detail in the following.
The relation between the abnormal regulation of CDK4 activity and carcinogenesis is observed in several cancer tissues. Deletion of p16 and p15 genes producing the proteins that inhibit CDK4 activity or overexpression of cyclin D1 that is essential for the CDK4 activity is observed in several kinds of cancer, which suggests that malignant phenotype may be induced when CDK4 activity is deregulated. Furthermore, it was reported that p16 knocked-out mouse has such a high carcinogenesis rate as p53 knocked-out mouse, which suggests that malfunction of p16 on CDK4 regulation is a cause of carcinogenesis. From these experimental results, deregulation of CDK4 activity may be a certain cause of carcinogenesis and play a role in maintenance of phenotype of cancer cell. Therefore, it is highly probable that CDK4 inhibitors have an anti-cancer effect.
It was reported that overexpression of cyclin E that is essential for CDK2 activity is observed in some breast cancers, is deeply associated with metastasis of breast cancer, inhibits cell apoptosis under low serum condition, and induces anchorage independent growth, and that hyperproliferation and neoplasia of mammary epithelial cells are observed in transformed animal where CDK2 is overexpressed using MMTV promoter. This strongly suggests that CDK2 activity is related with the progress or maintenance of cell transformation and CDK2 inhibitors may have an anti-cancer effect.
Furthermore, it has been gradually discovered that CDC2(CDK1), CDK3, CDK5, CDK6, CDK7, etc. play an important role in each phase of cell division. These are classified into CDKs family. In addition to cyclin D1 and E, cyclin A, B, C, D2, D3, D4, F and G are also classified into the same family.
On the basis of the above-mentioned researches, efficient inhibitors against these CDKs are recognized to be useful as an anti-cancer agent. Therefore, recently, some inhibitors have been developed.
As the effective CDKs inhibitor developed hitherto, Flavopiridol (EP 0,241,003 (1987) and 0,366,061 (1990)) represented by the following formula (2) can be mentioned: 
In addition, a purine derivative represented by the following formula (3): 
has been reported (WO 97/20842), and a compound represented by the following formula (4): 
having a quite different structure has been reported as an effective CDKs inhibitor (WO 98/33798).
However, the CDKs inhibitors developed up to now did not show satisfactory effects. Therefore, the present inventors have made extensive researches on CDKs inhibitors, particularly on flavone compounds and as a result, found that the above compound of formula (1) which has a quite different structure effectively inhibits the aforementioned CDKs and then, completed the present invention.
Therefore, the object of the present invention is to provide a novel 3-hydroxychromen-4-one derivative of formula (1), as defined above, pharmaceutically acceptable salt, hydrate, solvate or isomer thereof having an inhibitory activity for CDKs.
It is another object of the present invention to provide a process for preparing the compound of formula (1).
It is still another object of the present invention to provide a composition for suppression or treatment of cancer and diseases induced by cell proliferation such as inflammation, angiostenosis, angiogenesis, etc. comprising the compound of formula (1) as an active component together with pharmaceutically acceptable carriers.
In this specification, CDKs includes CDK2, CDK4, CDC2(CDK1), CDK3, CDK5, CDK6, CDK7, etc. and cyclins include cyclin D1, E, A, B, C, D2, D3, D4, F and G.
The present invention relates to a novel 3-hydroxychromen-4-one derivative represented by the following formula (1): 
in which
A represents hydrogen or nitro, or represents amino which is optionally substituted by C1-C4-alkylcarbonyl or carbamoyl, or represents a structure selected from a group consisting of 
wherein R4 represents hydrogen or C1-C6-alkyl which is optionally substituted by amino or hydroxy, R3 represents C1-C6-alkyl which is optionally substituted by amino or hydroxy and D represents halogen,
B represents methyl, or represents amino which is optionally mono- or disubstituted by substituents selected from a group consisting of C1-C6-alkyl, hydroxy-C1-C6-alkyl, C3-C6-cycloalkyl, acetyl, phenyl, benzyl and piperidinyl,
X, Y and Z independently of one another represent hydrogen, hydroxy, nitro, cyano or halogen, or represent amino which is optionally substituted by C1-C4-alkyl, C1-C4-alkylcarbonyl or carbamoyl, or represent C1-C4-alkyl which is optionally substituted by hydroxy or halogen,
pharmaceutically acceptable salt, hydrate, solvate or isomer thereof which exhibits a suppressive and therapeutic effect for cancer and diseases induced by cell proliferation such as inflammation, angiostenosis, angiogenesis, etc. by the inhibition of CDKs activities.
Since the compound of formula (1) according to the present invention may have asymmetric carbon atoms depending on the substituents, they can be present in the form of individual enantiomers or diastereomers, or mixtures thereof including racemates. Thus, the present invention also includes all of these isomers and their mixtures.
Also, the compound of formula (1) according to the present invention can form a pharmaceutically acceptable salt. Such salt includes non-toxic acid addition salt containing pharmaceutically acceptable anion, for example a salt with inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydriodic acid, etc., a salt with organic carboxylic acids such as tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trofluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, etc., or a salt with sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, etc.
Typical examples of the compound of formula (1) according to the present invention are
8-amino-2-(3 -amino-4-hydroxyphenyl)-3 -hydroxy-6-methyl-4H-chromen-4-one (Compound 1);
2-(4-hydroxy-3 -nitrophenyl)-3-hydroxy-6-methyl-8-nitro-4H-chromen-4-one (Compound 2);
N-[2-(4-hydroxy-3-nitrophenyl)-3-hydroxy-6-methyl-4-oxo-4H-chromen-8-yl]acetamide (Compound 3);
8-amino-2-(4-hydroxyphenyl)-3-hydroxy-6-methyl-4H-chromen-4-one (Compound 4);
2-(3 -amino-4-hydroxyphenyl)-3-hydroxy-6-methyl-4H-chromen-4-one (Compound 5);
N-[2-hydroxy-5-(3-hydroxy-6-methyl-4-oxo-4H-chromen-2-yl)phenyl]acetamide (Compound 6);
N-{2-[4-hydroxy-3-(isopropylamino)phenyl]-3-hydroxy-6-methyl-4-oxo-4H-chromen-8-yl}acetamide (Compound 7A);
N-[2-(3-amino-4-hydroxyphenyl)-3-hydroxy-6-methyl-4-oxo-4H-chromen-8-yl]acetamide (Compound 7B);
2-(3-fluoro-4-hydroxyphenyl)-3-hydroxy-6-methyl-8-nitro-4H-chromen-4-one (Compound 8);
8-amino-2-(3-fluoro-4-hydroxyphenyl)-3-hydroxy-6-methyl-4H-chromen-4-one (Compound 9);
N-{5-[8-(ureido)-3-hydroxy-6-methyl-4-oxo-4H-chromen-2-yl]-2-hydroxyphenyl}urea (Compound 10);
8-amino-2-(4-aminophenyl)-3-hydroxy-6-methyl-4H-chromen-4-one (Compound 11);
8-amino-3 -hydroxy-2-(3-hydroxyphenyl)-6-methyl-4H-chromen-4-one (Compound 12);
3-hydroxy-6-methyl-2-(2,3,4-trihydroxyphenyl)-4H-chromen-4-one (Compound 13);
2-(2-bromo-3,4-dihydroxyphenyl)-3-hydroxy-6-methyl-4H-chromen-4-one (Compound 14);
2-[3-hydroxy-2-(4-hydroxyphenyl)-6-methyl-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 15);
2-[2-(3-fluoro-4-hydroxyphenyl)-3-hydroxy-6-methyl-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 16);
2-[2-(3-chloro-4-hydroxyphenyl)-3-hydroxy-6-methyl-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 17);
2-[2-(3-bromo-4-hydroxyphenyl)-3-hydroxy-6-methyl-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 18);
5-[8-(1,1-dioxo-1xcex6-isothiazolidin-2-yl)-3-hydroxy-6-methyl-4-oxo-4H-chromen-2-yl]-2-hydroxybenzonitrile (Compound 19);
2-[2-(2,4-dihydroxyphenyl)-3-hydroxy-6-methyl-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 20);
2-[2-(3-chloro-4-fluorophenyl)-3-hydroxy-6-methyl-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 21);
2-[3-hydroxy-2-(4-hydroxy-3-methylphenyl)-6-methyl-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 22);
2-{3-hydroxy-2-[4-hydroxy-3-(hydroxymethyl)phenyl]-6-methyl-4-oxo-4H-chromen-8-yl}-1xcex6-isothiazolidin-1,1-dione (Compound 23);
2-[3-hydroxy-2-(4-hydroxy-3-trifluoromethylphenyl)-6-methyl-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 24);
2-[6-amino-3-hydroxy-2-(4-hydroxyphenyl)-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 25);
2-[6-(dimethylamino)-3-hydroxy-2-(4-hydroxyphenyl)-4-oxo-4H-chromen-8-y]-1xcex6-isothiazolidin-1,1-dione (Compound 26);
2-[6-(diethylamino)-3-hydroxy-2-(4-hydroxyphenyl)-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 27);
2-[6-(benzylamino)-3-hydroxy-2-(4-hydroxyphenyl)-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 28);
2-[3 -hydroxy-2-(4-hydroxyphenyl)-4-oxo-6-(4-piperidinylamino)-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 29);
2-[6-(cyclohexylamino)-3-hydroxy-2-(4-hydroxyphenyl)-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 30);
2-[6-anilino-3-hydroxy-2-(4-hydroxyphenyl)-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 31);
2-[3-hydroxy-2-(4-hydroxyphenyl)-6-(methylamino)-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 32);
2-{3-hydroxy-6-[(2-hydroxyethyl)(methyl)amino]-2-(4-hydroxyphenyl)-4-oxo-4H-chromen-8-yl}-1xcex6-isothiazolidin-1,1-dione (Compound 33);
N-[2-(3-chloro-4-hydroxyphenyl)-8-(1,1dioxo-1xcex6-isothiazolidin-2-yl)-3-hydroxy-4-oxo-4H-chromen-6-yl]acetamide (Compound 34);
2-[6-amino-2-(3-chloro-4-hydroxyphenyl)-3-hydroxy-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 35);
2-[2-(3-chloro-4-hydroxyphenyl)-6-(dimethylamino)-3-hydroxy-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 36);
2-[2-(3-chloro-4-hydroxyphenyl)-6-(methylamino)-3-hydroxy-4-oxo-4H-chromen-8-yl]-1xcex6-isothiazolidin-1,1-dione (Compound 37);
2-(3-chloro-4-hydroxyphenyl)-3-hydroxy-6-methyl-8-(1-methyl-4-piperidinyl)-4H-chromen-4-one (Compound 38);
2-(3-chloro-4-hydroxyphenyl)-3-hydroxy-6-methyl-8-(4-pyridinyl)-4H-chromen-4-one (Compound 39);
4-[2-(3-chloro-4-hydroxyphenyl)-3-hydroxy-6-methyl-4-oxo-4H-chromen-8-yl]-1-methylpyridinium bromide (Compound 40);
2-(4-hydroxyphenyl)-3-hydroxy-6-methyl-8-(1-methyl-4-piperidinyl)-4H-chromen-4-one (Compound 41);
3-hydroxy-2-(4-hydroxy-3-methylphenyl)-6-methyl-8-(1-methyl-4-piperidinyl)-4H-chromen-4-one (Compound 42);
3 -hydroxy-2-(4-hydroxy-3-trifluoromethylphenyl)-6-methyl-8-(1-methyl-4-piperidinyl)-4H-chromen-4-one (Compound 43);
2-(3-chloro-4-hydroxyphenyl)-3-hydroxy-8-[1-(2-hydroxyethyl)-4-piperidinyl]-6-methyl-4H-chromen-4-one (Compound 44); and
8-[1-(2-aminoethyl)-4-piperidinyl]-2-(3-chloro-4-hydroxyphenyl)-3-hydroxy-6-methyl-4H-chromen-4-one (Compound 45).
The compound of formula (1) of the present invention may be prepared by a process as described in the following and thus, it is another object of the present invention to provide such a process.
The compound of formula (1) of the present invention may be prepared characterized in that
(a) a compound represented by the following formula (5): 
in which A and B are defined as previously described, is reacted with an aldehyde represented by the following formula (6): 
in which X, Y and Z are defined as previously described, to produce a compound represented by the following formula (7): 
in which A, B, X, Y and Z are defined as previously described, and the compound of formula (7) thus prepared is cyclized in the presence of a base to produce the compound of formula (1);
(b) a compound represented by the following formula (8): 
in which B, X, Y and Z are defined as previously described and P represents hydroxy-protecting group, preferably methyl or benzyl, is reacted with 3-chloropropanesulfonyl-chloride in the presence of a base and a catalyst to produce a compound represented by the following formula (9): 
in which B, X, Y, Z and P are defined as previously described, and the compound of formula (9) thus prepared is deprotected to produce a compound represented by the following formula (1a): 
in which B, X, Y and Z are defined as previously described;
(c) a compound represented by the following formula (10): 
in which B and P are defined as previously described, L represents leaving group, preferably halogen, and Xxe2x80x2, Yxe2x80x2 and Zxe2x80x2 each are identical with X, Y and Z, respectively, but hydroxy group(s) is(are) protected, is reacted with 4-halogenopyridine in the presence of a base and a catalyst and then deprotected to produce a compound represented by the following formula (1b): 
in which B, X, Y and Z are defined as previously described;
(d) the compound obtained before the deprotection step in process variant (c) is reacted with a compound represented by the following formula (11):
R3Dxe2x80x83xe2x80x83(11). 
in which R3 and D are defined as previously described, and then deprotected to produce a compound represented by the following formula (1c): 
in which B, X, Y, Z, R3 and D are defined as previously described;
(e) the compound obtained before the deprotection step in process variant (d) is reduced and deprotected to produce a compound represented by the following formula (1d): 
in which B, X, Y and Z are defined as previously described and R4 is identical with R4 but other than hydrogen;
or further hydrolysis protection, deprotection, reduction or amidation reaction may be carried out.
Hereinafter, the process according to the present invention is more specifically explained.
In process variant (a), the 2-hydroxyacetophenone derivative of formula (5) is reacted with 3 equivalents of the benzaldehyde derivative of formula (6) based on the compound of formula (5) to produce the compound of formula (7). As the base, 3 equivalents of sodium hydroxide based on the compound of formula (5) are preferably used. The reaction is preferably carried out in 80% aqueous ethanol solution for 3 hours at room temperature. Subsequently, the compound of formula (7) thus obtained is reacted in methanol solvent in the presence of excess 10% aqueous sodium hydroxide solution and excess hydrogen peroxide for 2 hours at room temperature to produce the compound of formula (1).
In process variant (b), the compound of formula (8) is dissolved in a solvent such as dichloromethane, reacted with 3-chloropropanesulfonylchloride in the presence of triethylamine and catalytic amount of dimethylaminopyridine at room temperature, and then concentrated. The resulting residue is dissolved again in dimethylformamide solvent and aqueous sodium hydroxide solution is added thereto. The mixture thus obtained is reacted for 30 minutes at 50xc2x0 C. to produce the compound of formula (9). The compound of formula (9) is then reduced with hydrogen gas under a solvent system such as methanol/dichloromethane, or reacted with borontribromide in a solvent such as dichloromethane to produce the compound of formula (1a).
The compound of formula (8) used as a starting material in process variant (b) may be prepared according to the method as depicted in the following reaction scheme 1: 
in which
B, X, Y, Z and P are defined as previously described, and
R1 and R2 independently of one another represent hydrogen, oxo or acyl group, but both of them are not hydrogen.
In the reaction scheme 1, the process for preparing the compound of formula (15) from the compound of formula (12) may be carried out according to the same procedure as process variant (a). The compound of formula (15) thus prepared is refluxed with iodomethane or benzyl bromide in acetone solvent in the presence of potassium carbonate to produce the compound of formula (16) wherein 3-hydroxy group on the chromene ring is protected by a group of methyl or benzyl. The compound of formula (16) wherein 8-position of the chromene ring is substituted by nitro group is reduced by hydrogen gas. Otherwise, when the same position in the compound of formula (16) is substituted by acylamino group, BOC group is introduced into the amide group and the resulting compound is hydrolyzed to produce the compound of formula (8).
Process variants (c), (d) and (e) may be explained as follows. The starting compound of formula (10) is heated with 1.5 equivalent of bispinacolatodiboron, 3 equivalents of potassium acetate and 5 mol % of dichlorobistriphenylphosphine palladium in N,N-dimethylformamide solvent under nitrogen atmosphere to 80 to 90xc2x0 C. and reacted for 2 hours. The reactants are cooled to room temperature, reacted with 2 equivalents of 4-bromopyridine hydrochloride, 5 mol % of dichlorobistriphenylphosphine palladium and 5 equivalents of 2M aqueous sodium carbonate solution for 15 hours, and then deprotected to produce the compound of formula (1b). The compound obtained before deprotection step to the formula (1b) is heated under reflux with 2 equivalents of the compound of formula (11) in acetone or acetonitrile solvent for 3 hours and then deprotected to produce the compound of formula (1c). Also, the compound obtained before deprotection step to the formula (1c) is dissolved in 50% methanol/dichloromethane, reacted with 5 mol % of platinum oxide under room temperature and 1 atm of hydrogen atmosphere for 48 hours, and then deprotected to produce the compound of formula (1d). In process variants (c), (d) and (e), the deprotection step is carried out by adding the compound to dry dichloromethane, adding 5 equivalents of borontribrornide thereto, and reacting for 10 hours at room temperature.
Further, the compound of formula (10) used as a starting material in process variants (c), (d) and (e) may be prepared according to the method as depicted in the following reaction scheme 2: 
in which
B, L, X Y, Z, P, Xxe2x80x2, Yxe2x80x2 and Zxe2x80x2 are defined as previously described.
In reaction scheme 2, the compound of formula (17) is reacted with the benzaldehyde of formula (13) and 3 equivalents of sodium hydroxide in the solvent of 80% aqueous ethanol solution for 17 hours at room temperature to produce the compound of formula (18). This compound is reacted with 3 equivalents of 10% aqueous sodium hydroxide solution and 3 equivalents of hydrogen peroxide in methanol solvent for 3 hours at room temperature to produce the compound of formula (19). The compound of formula (19) thus obtained is reacted with 2 equivalents of iodomethane and potassium carbonate in acetone solvent for 30 minutes at room temperature to produce the compound of formula (10).
The reaction conditions including reaction solvent, base, amount of reactants, etc. in the process according to the present invention are not restricted to those as mentioned above and can easily be selected by optionally combining the various synthetic ways described in the present specification or known in the art. And such a combination may be easily carried out by one of ordinary skill in the art. The following preparations and examples may also be referred to the specific reaction conditions of the above process.
After the reaction is completed, the product may be isolated and purified by conventional work up processes such as chromatography, recrystallization, etc.
The compound of formula (1) of the present invention can be effectively used as medicines for suppression or treatment of cancer and diseases induced by cell proliferation such as inflammation, angiostenosis, angiogenesis, etc. due to its excellent inhibitory activity against CDKs. Therefore, another object of the present invention is to provide a composition for suppression or treatment of cancer and diseases induced by cell proliferation such as inflammation, angiostenosis, angiogenesis, etc. characterized by comprising the compound of formula (1), pharmaceutically acceptable salt, hydrate, solvate or isomer thereof as an active component together with pharmaceutically acceptable carriers.
When the active compound according to the present invention is used for clinical purpose, it is preferably administered to the subject patient in an amount ranging from 1 to 50 mg per kg of body weight a day. The total daily dosage may be administered in one time or over several times. However, the specific administration dosage for the specific patient can be varied with the specific compound used, body weight of the subject patient, sex, hygienic condition, diet, time or method of administration, excretion rate, mixing ratio of the agent, severity of the disease to be treated, etc.
The compound of the present invention may be administered in the form of injections or oral preparations.
Injections, for example, sterilized aqueous or oily suspension for injection, can be prepared according to the known procedure using suitable dispersing agent, wetting agent, or suspending agent. Solvents that can be used for preparing injections include water, Ringer""s fluid and isotonic NaCl solution, and also sterilized fixing oil may be conveniently used as the solvent or suspending media. Any non-stimulative fixing oil including mono-, di-glyceride may be used for this purpose. Fatty acid such as oleic acid may also be used for injections.
As the solid preparation for oral administration, capsules, tablets, pills, powders and granules, etc., preferably capsules and tablets can be mentioned. It is also desirable for tablets and pills to be formulated into enteric-coated preparation. The solid preparations may be prepared by mixing the active compound of formula (1) according to the present invention with at least one carrier selected from a group consisting of inactive diluents such as sucrose, lactose, starch, etc., lubricants such as magnesium stearate, disintegrating agent and binding agent.
When the compound of the present invention is clinically administered for the purpose of treating cancer, the active compound of formula (1) may be simultaneously administered with one or more selected from the known anti-cancer agents. As the anti-cancer agent that can be administered together with the compound of the present invention, 5-fluorouracil, cisplatin, doxorubicin, taxol, Gemcitabine, etc. can be mentioned.
However, anti-cancer preparations containing the compound of the present invention are not limited to those as explained above, and any agent capable of preventing or treating cancer may be included.
The present invention will be more specifically explained in the following examples and experiments. However, it should be understand that these examples and experiments are intended to illustrate the present invention but not in any manner to limit the scope of the present invention.
Preparation 1
Synthesis of (E)-3-[4-(benzyloxy)-3-nitrophenyl]-1-(2-hydroxy-5-methyl-3-nitrophenyl)-2-propen-1-one
2-hydroxy-5-methyl-3-nitroacetophenone (200 mg, 1.02 mmol), 4-benzyloxy-3-nitrobenzaldehyde (300 mg, 1.16 mmol) and 3 equivalents of sodium hydroxide (120 mg) were introduced into 80% aqueous ethanol solution and the resulting mixture was stirred for 3 hours at room temperature. The reaction solution was acidified by 2N hydrochloric acid solution and diluted with water. The resulting solid was filtered, washed with large amount of water and methanol, and then dried to give 430 mg (Yield 97%) of the title compound as a yellow solid.
1H NMR (CDCl3, ppm); xcex48.06-7.75 (5H, m), 7.60-7.15 (7H, m), 5.31 (2H, s), 2.43 (3H, s)
FAB MS(m/e)=435[M++1]
Preparation 2
Synthesis of 2-[4-(benzyloxy)-3-nitrophenyl]-3-hydroxy-6-methyl-8-nitro-4H-chromen-4-one
The compound prepared in Preparation 1 (400 mg, 0.92 mmol) was introduced into methanol, and aqueous hydrogen peroxide (0.5 ml) and 10% aqueous sodium hydroxide solution (0.5 ml) were added thereto at room temperature. The resulting mixture was stirred for 2 hours at room temperature, acidified by 2N hydrochloric acid solution, and diluted with water. The solid having a pale yellow color thus obtained was filtered, washed with large amount of water and methanol, and then dried to give 200 mg (Yield 48.5%) of the title compound.
1H NMR (CDCl3, ppm); xcex48.84 (2H, s), 8.44 (1H, s), 8.19 (2H, d), 7.34-7.17 (5H, m)
FAB MS(m/e)=449[M++1]